ChIP-Atlas: SRX13166188

Antigen

Cell type Class: Neural
Cell type: Hippocampus
MeSH Description: A curved elevation of GRAY MATTER extending the entire length of the floor of the TEMPORAL HORN of the LATERAL VENTRICLE (see also TEMPORAL LOBE). The hippocampus proper, subiculum, and DENTATE GYRUS constitute the hippocampal formation. Sometimes authors include the ENTORHINAL CORTEX in the hippocampal formation.

library_name: GSM5694683
library_strategy: ChIP-Seq
library_source: GENOMIC
library_selection: ChIP
library_construction_protocol: virally transduced (shFoxg1, shNeurod1 or shLuciferase) primary hippocampal neurons were fixed and collected for subsequent ChIP-seq experiments according to the instructions of the servicing company (Active Motif). Briefly, cells were fixed by adding 1/10 volume formaldehyde solution (11% Formaldehyde, 0.1 M NaCl, 1 mM EDTA pH=8.0, 50 mM HEPES pH=7.9) directly to the medium and incubated at room temperature (RT) with agitation. Fixation was stopped by adding 1/20 volume 2.5 M Glycine solution (Roth, Germany) and incubating for 5 min at RT. Cells were scraped from the plates and transferred into conical tubes, and centrifuged at 800 rpm at 4°C for 10 min. Cells were resuspended and washed twice in chilled PBS-Igepal solution (0.5% Igepal CA-360 (Sigma) in 1X PBS pH 7.4 (Gibco)), and a third time in PBS-Igepal-PMSF (phenylmethylsulfonyl fluoride) solution (0.5% Igepal, 1 mM PMSF (Sigma) in 1X PBS pH 7.4). The supernatant was completely removed and cell pellets were snap frozen in liquid nitrogen and stored at -80°C until following ChIP-seq experiments. ChIP-Seq libraries were generated from the ChIP-DNA using a custom Illumina library type on an automated system (Apollo 342, Wafergen Biosystems/Takara) ChIP-Seq libraries were sequenced on Illumina NextSeq 500 as 75-nt single end reads.